Structure and function of renin and prorenin.

肾素和肾素原的结构和功能。

• 批准号: 01060003
• 负责人: MURAKAMI Kazuo
• 金额: $ 133.12万
• 依托单位: University of Tsukuba
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Specially Promoted Research
• 财政年份: 1989
• 资助国家: 日本
• 起止时间: 1989 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01060003/
• 关键词: Renin; Prorenin; Angiotensinogen; X ray analysis; Embryonic Engineering; Processing; Transgenic mice; Hypertension; 高血圧マウス; 遺伝子工学; プロテイン・エンジニアリング; 組織特異的発現

1) X ray structure of reninThree-dimensional structure of mouse submaxillary renin and recombinant human renin have been determined at 2.5A^^ﾟ - 3.0A^^ﾟ resolution by X-ray analysis.2) Processing enzymesThe cDNA of prorenin processing enzyme in mouse submaxillary glands was cloned and its complete nucleotide sequence was determined, identify that prorenin processing enzyme is identical with proEGF processing enzyme. The processing enzyme processed the submaxillary prorenin to renin, but not kidney prorenin suggesting its organ specificity.Several candidates (yeast Kex2 like processing enzyme) for kidney prorenin processing enzyme or for prohormones and proenzymes have been identified and their complete amino acid sequences have been determined. These results suggests that not only prorenin but also prohormones and proenzymes are processed by substrate- or organ-specific manner.3) Gene regulation of renin and angiotensinogen in vivoHuman renin and human angiotensinogen genes both separately and in concert have been introduced into mice by cross-mating.Strains carring both the human genes demonstrated a chronically sustained increase in blood pressure. Thus, we have established new transgenic mice (Tsukuba Hypertensive Mice) as the first model of human high renin hypertension.

1)2）加工酶克隆了小鼠颌下腺中原肾素加工酶的cDNA，并测定了其全序列，鉴定了原肾素加工酶与原EGF加工酶的同源性。该加工酶将颌下前肾素加工成肾素，但不将肾前肾素加工成肾素，这表明其具有器官特异性。已鉴定出肾前肾素加工酶或激素原和前酶的几种候选酶（酵母Kex 2样加工酶），并确定了它们的完整氨基酸序列。这些结果表明，不仅是原肾素，而且激素原和酶原也是通过底物或器官特异性方式加工的。3）体内肾素和血管紧张素原的基因调控通过交叉交配将人肾素和人血管紧张素原基因分别或同时导入小鼠，携带这两种人类基因的菌株表现出慢性持续的血压升高。因此，我们建立了新的转基因小鼠（筑波高血压小鼠）作为人类高肾素高血压的第一个模型。

项目成果

Toshihisa HATAE: "Expression and purification of human angiotensinogen in Chinese hamster ovary cells." Biochim.Biophys.Acta. 1121. 335-338 (1992)

Toshihisa HATAE：“人血管紧张素原在中国仓鼠卵巢细胞中的表达和纯化。”

Akiyoshi FUKAMIZU: "Structure and expression of the human angiotensinogen gene. Identification of a unique and highly active promoter." J.Biol. Chem.265. 7576-7582 (1990)

Akiyoshi FUKAMIZU：“人类血管紧张素原基因的结构和表达。独特且高活性启动子的鉴定。”

深水 昭吉,村上 和雄: "実験医学" (株)羊土社, 7 (1990)

深见明吉、村上一夫：《实验医学》Yodosha Co., Ltd.，7 (1990)

宮崎 均、近藤 元宏、村上 和雄: "ホルモンと臨床" (有)医学の世界社, 6 (1991)

宫崎仁、近藤元博、村上一夫：“激素与临床实践” Igaku no Sekaisha，6 (1991)

Takeshi Yamauchi,ea al.: "Siteーdirected mutagenesis of human prorenin.Substitution of three arginine residues in the propeptide with glutamine residues yeilds active prorenin" Journal of Biochemistry. 107. 27-30 (1990)

Takeshi Yamauchi等人：“人肾素原的定点诱变。用谷氨酰胺残基取代前肽中的三个精氨酸残基产生活性肾素原”《生物化学杂志》107. 27-30 (1990)。